This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We are using a specially designed PCR template for homologous replacement of the endogenous protein promoters in yeast to inducible promoters. Expressed several protein substrates of endogenous kinases from these inducible promoters and in order to map phosphorylation sites on the proteins. We are testing the technique by mapping phosphorylation sites on several of these proteins including mcm3, bud3, boi2, lte1, ned1.